Sodium-Calcium Exchange in Intracellular Calcium Handling of Human Airway Smooth Muscle 英文参考文献.docVIP

Sodium-CalciumExchangeinIntracellularCalcium HandlingofHumanAirwaySmoothMuscle VenkatachalemSathish1,2,PhilippeF.Delmotte1,MichaelA.Thompson2,ChristinaM.Pabelick1,2,GaryC. Sieck1,2,Y.S.Prakash1,2 * 1Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota, United States of America, 2Department of Anesthesiology, Mayo Clinic, Rochester,Minnesota,UnitedStatesofAmerica Abstract Enhancedairwaycontractilityfollowinginflammationbycytokinessuchastumornecrosisfactoralpha(TNFa)orinterleukin- 13(IL-13)involvesincreasedintracellular Ca ([Ca ]i)levelsinairway smoothmuscle(ASM).InASM,plasma membrane Ca 2+ 2+ 2+ fluxes form a key component of [Ca2+]i regulation. There is now growing evidence that the bidirectional plasma + 2+ 2+ membrane Na /Ca exchanger (NCX) contributes to ASM [Ca ]i regulation. In the present study, we examined NCX expressionandfunctioninhumanASMcellsundernormalconditions,andfollowingexposuretoTNFaorIL-13.Western blot analysis showed significant expression of the NCX1 isoform, with increased NCX1 levels by both cytokines, effects blunted by inhibitors of nuclear factor NF-kB or mitogen-activated protein kinase. Cytokine-mediated increase in NCX1 involvedenhancedtranscriptionfollowedbyproteinsynthesis.NCX2andNCX3remainedundetectableevenincytokine- stimulated ASM. In fura-2 loaded human ASM cells, NCX-mediated inward Ca exchange as well as outward exchange (measuredasratesofchangein[Ca ]i)waselicitedbyalteringextracellularNa andCa levels.ContributionofNCXwas verified by measuring [Na ]i using the fluorescent Na indicator SBFI. NCX-mediated inward exchange was verified by demonstratingpreventionofrising[Ca ]iorfalling[Na+]iinthepresenceoftheNCXinhibitorKBR7943.Inwardexchange- mode NCX was increased by both TNFa and IL-13 to a greater extent than outward exchange. NCX siRNA transfection substantially blunted outward exchange and inward exchange modes. Finally, inhibition